WO2015170336A1 - 16α-HETEROARYL PREGNENOLONE ACETATE AND A PROCESS FOR PREPARATION THEREOF - Google Patents
16α-HETEROARYL PREGNENOLONE ACETATE AND A PROCESS FOR PREPARATION THEREOF Download PDFInfo
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- WO2015170336A1 WO2015170336A1 PCT/IN2015/000188 IN2015000188W WO2015170336A1 WO 2015170336 A1 WO2015170336 A1 WO 2015170336A1 IN 2015000188 W IN2015000188 W IN 2015000188W WO 2015170336 A1 WO2015170336 A1 WO 2015170336A1
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- compound
- cell line
- cancer cell
- pregnenolone acetate
- benzimidazole
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- 238000002360 preparation method Methods 0.000 title claims description 23
- 238000000034 method Methods 0.000 title claims description 20
- CRRKVZVYZQXICQ-RJJCNJEVSA-N Pregnenolone acetate Chemical compound C([C@@H]12)C[C@]3(C)[C@@H](C(C)=O)CC[C@H]3[C@@H]1CC=C1[C@]2(C)CC[C@H](OC(=O)C)C1 CRRKVZVYZQXICQ-RJJCNJEVSA-N 0.000 title description 5
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 claims abstract description 53
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 31
- 201000011510 cancer Diseases 0.000 claims abstract description 30
- HYZJCKYKOHLVJF-UHFFFAOYSA-N 1H-benzimidazole Chemical compound C1=CC=C2NC=NC2=C1 HYZJCKYKOHLVJF-UHFFFAOYSA-N 0.000 claims abstract description 21
- 230000001472 cytotoxic effect Effects 0.000 claims abstract description 15
- 229940125904 compound 1 Drugs 0.000 claims abstract description 13
- 206010006187 Breast cancer Diseases 0.000 claims abstract description 12
- 208000026310 Breast neoplasm Diseases 0.000 claims abstract description 12
- 210000002307 prostate Anatomy 0.000 claims abstract description 12
- 150000002576 ketones Chemical class 0.000 claims abstract description 11
- 238000000338 in vitro Methods 0.000 claims abstract description 8
- 238000006845 Michael addition reaction Methods 0.000 claims abstract description 6
- 150000001875 compounds Chemical class 0.000 claims description 48
- MZWRIOUCMXPLKV-RFOVXIPZSA-N 16-Dehydropregnenolone acetate Chemical compound C([C@@H]12)C[C@]3(C)C(C(C)=O)=CC[C@H]3[C@@H]1CC=C1[C@]2(C)CC[C@H](OC(=O)C)C1 MZWRIOUCMXPLKV-RFOVXIPZSA-N 0.000 claims description 30
- YLFRRPUBVUAHSR-UHFFFAOYSA-N 16-dehydro-pregnenolone Natural products C1C=C2CC(O)CCC2(C)C2C1C1CC=C(C(=O)C)C1(C)CC2 YLFRRPUBVUAHSR-UHFFFAOYSA-N 0.000 claims description 26
- 239000012043 crude product Substances 0.000 claims description 17
- OAYLNYINCPYISS-UHFFFAOYSA-N ethyl acetate;hexane Chemical compound CCCCCC.CCOC(C)=O OAYLNYINCPYISS-UHFFFAOYSA-N 0.000 claims description 17
- 239000004570 mortar (masonry) Substances 0.000 claims description 16
- 238000010898 silica gel chromatography Methods 0.000 claims description 16
- 239000002904 solvent Substances 0.000 claims description 6
- 229940125782 compound 2 Drugs 0.000 claims description 5
- 230000001678 irradiating effect Effects 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 230000003637 steroidlike Effects 0.000 abstract description 26
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 abstract description 16
- 229960004679 doxorubicin Drugs 0.000 abstract description 8
- 239000003814 drug Substances 0.000 abstract description 6
- 229940079593 drug Drugs 0.000 abstract description 5
- 210000004027 cell Anatomy 0.000 description 41
- 238000006243 chemical reaction Methods 0.000 description 17
- 239000007787 solid Substances 0.000 description 16
- 239000003480 eluent Substances 0.000 description 11
- 150000003431 steroids Chemical class 0.000 description 9
- 125000000623 heterocyclic group Chemical group 0.000 description 6
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 6
- 238000003786 synthesis reaction Methods 0.000 description 6
- 238000005481 NMR spectroscopy Methods 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 5
- 239000003054 catalyst Substances 0.000 description 5
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 4
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 4
- 230000003013 cytotoxicity Effects 0.000 description 4
- 231100000135 cytotoxicity Toxicity 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 3
- 206010060862 Prostate cancer Diseases 0.000 description 3
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 3
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 3
- 230000001093 anti-cancer Effects 0.000 description 3
- 125000003785 benzimidazolyl group Chemical group N1=C(NC2=C1C=CC=C2)* 0.000 description 3
- 230000004071 biological effect Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 125000002883 imidazolyl group Chemical group 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 230000003389 potentiating effect Effects 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 125000001424 substituent group Chemical group 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical group C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- 102000001307 androgen receptors Human genes 0.000 description 2
- 108010080146 androgen receptors Proteins 0.000 description 2
- SCWLBXZTXMYTLF-UHFFFAOYSA-N cyclopropane-1,2-dicarbohydrazide Chemical compound NNC(=O)C1CC1C(=O)NN SCWLBXZTXMYTLF-UHFFFAOYSA-N 0.000 description 2
- 231100000433 cytotoxic Toxicity 0.000 description 2
- 235000019439 ethyl acetate Nutrition 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 238000010348 incorporation Methods 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- ONBQEOIKXPHGMB-VBSBHUPXSA-N 1-[2-[(2s,3r,4s,5r)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]oxy-4,6-dihydroxyphenyl]-3-(4-hydroxyphenyl)propan-1-one Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1OC1=CC(O)=CC(O)=C1C(=O)CCC1=CC=C(O)C=C1 ONBQEOIKXPHGMB-VBSBHUPXSA-N 0.000 description 1
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- AZKSAVLVSZKNRD-UHFFFAOYSA-M 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide Chemical compound [Br-].S1C(C)=C(C)N=C1[N+]1=NC(C=2C=CC=CC=2)=NN1C1=CC=CC=C1 AZKSAVLVSZKNRD-UHFFFAOYSA-M 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 229940127512 Androgen Synthesis Inhibitors Drugs 0.000 description 1
- 102000014654 Aromatase Human genes 0.000 description 1
- 108010078554 Aromatase Proteins 0.000 description 1
- OSMCIRLKYVPBKF-MQFHIODPSA-N C[C@](CC1)([C@@H](C[C@H]2[n]3c(cccc4)c4nc3)[C@H]3[C@H]1[C@@](C)(CC[C@@H](C1)OC(C)=O)C1=CC3)[C@H]2C(C)=O Chemical compound C[C@](CC1)([C@@H](C[C@H]2[n]3c(cccc4)c4nc3)[C@H]3[C@H]1[C@@](C)(CC[C@@H](C1)OC(C)=O)C1=CC3)[C@H]2C(C)=O OSMCIRLKYVPBKF-MQFHIODPSA-N 0.000 description 1
- CXMSNDUDBRCABN-SQLUACQTSA-N C[C@](CC1)([C@@H](C[C@H]2[n]3cncc3)[C@H]3[C@H]1[C@@](C)(CC[C@@H](C1)OC(C)=O)C1=CC3)[C@H]2C(C)=O Chemical compound C[C@](CC1)([C@@H](C[C@H]2[n]3cncc3)[C@H]3[C@H]1[C@@](C)(CC[C@@H](C1)OC(C)=O)C1=CC3)[C@H]2C(C)=O CXMSNDUDBRCABN-SQLUACQTSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 239000002841 Lewis acid Substances 0.000 description 1
- 231100000002 MTT assay Toxicity 0.000 description 1
- 238000000134 MTT assay Methods 0.000 description 1
- GZOSMCIZMLWJML-VJLLXTKPSA-N abiraterone Chemical compound C([C@H]1[C@H]2[C@@H]([C@]3(CC[C@H](O)CC3=CC2)C)CC[C@@]11C)C=C1C1=CC=CN=C1 GZOSMCIZMLWJML-VJLLXTKPSA-N 0.000 description 1
- 229960000853 abiraterone Drugs 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000007259 addition reaction Methods 0.000 description 1
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 1
- 239000003098 androgen Substances 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000000843 anti-fungal effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000002365 anti-tubercular Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 229940121375 antifungal agent Drugs 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 238000012925 biological evaluation Methods 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 150000001728 carbonyl compounds Chemical class 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229940126142 compound 16 Drugs 0.000 description 1
- 229940127089 cytotoxic agent Drugs 0.000 description 1
- 239000002254 cytotoxic agent Substances 0.000 description 1
- 231100000599 cytotoxic agent Toxicity 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 125000005331 diazinyl group Chemical group N1=NC(=CC=C1)* 0.000 description 1
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 1
- 231100001261 hazardous Toxicity 0.000 description 1
- -1 hetero steroids Chemical class 0.000 description 1
- 150000002460 imidazoles Chemical class 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- QNXSIUBBGPHDDE-UHFFFAOYSA-N indan-1-one Chemical class C1=CC=C2C(=O)CCC2=C1 QNXSIUBBGPHDDE-UHFFFAOYSA-N 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 150000007517 lewis acids Chemical class 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000006053 organic reaction Methods 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- AICOOMRHRUFYCM-ZRRPKQBOSA-N oxazine, 1 Chemical compound C([C@@H]1[C@H](C(C[C@]2(C)[C@@H]([C@H](C)N(C)C)[C@H](O)C[C@]21C)=O)CC1=CC2)C[C@H]1[C@@]1(C)[C@H]2N=C(C(C)C)OC1 AICOOMRHRUFYCM-ZRRPKQBOSA-N 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 238000009522 phase III clinical trial Methods 0.000 description 1
- 239000003586 protic polar solvent Substances 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 230000002381 testicular Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 125000003831 tetrazolyl group Chemical group 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 238000010626 work up procedure Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
- C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Definitions
- the present invention relates to 16a-heteroayl-l -yl pregnenolone acetate and a process for its preparation.
- the present invention particularly relates to compounds of 16cc-(lH-benzimidazol- l -yl)-pregnenolone acetate (1) and 16ct-(lH-imidazol- l -yl)- pregnenolone acetate (2).
- the present invention also relates to a process for the preparation of said compounds froml6-dehydropregnenolone acetate (16-DPA). More particularly the present invention relates to the anticancerous activity of the said compounds (1 and 2).
- Benzimidazole, imidazole and their derivatives are very important pharmacophores in medicinal chemistry showing a wide range of biological activities like antibacterial, antifungal, anticancer, antiviral, antituberculosis and antiinflammatory and antiageing agents.
- the main object of the present invention is to provide 16a-heteroayl- l -yl pregnenolone acetate and a process for preparation of the compounds.
- Another object of the present invention is to provide compounds 16a-( lH- benzimidazol-l -yl)-pregnenolone acetate (1) and 16ec-(lH-imidazol-l -yl)- pregnenolone acetate (2).
- Another object of the present invention is to provide a process for the preparation of 16a-(lH-benzimidazol-l -yl)-pregnenolone acetate (1) and 16a-(lH- imidazol-l -yl)-pregnenolone acetate (2) from 16-dehydropregnenolone acetate ( 16- DPA) without using solvent and catalyst under microwave irradiation.
- Still another objective of the present invention is to evaluate the in vitro cytotoxic activities of compound 1 and 2 against cervical HeLa cancer cell line, prostate DU 205 cancer cell line and breast cancer MCF-7 cell line in comparison to the drug doxorubicin.
- step b) irradiating the mixture obtained in step a) in a microwave reactor at a frequency of 2450 MHz for a time period ranging between 10 to 30 minutes,
- step b) purifying the crude product obtained in step b) by silica gel column chromatography using EtOAc-hexane taken in the ratio of 1 :4.
- step b) silica gel column chromatography using EtOAc-hexane taken in the ratio of 1 :4.
- Figure 1 shows the synthesis of compounds 16a-(lH-benzimidazol-l -yl)- pregnenolone acetate (1) and 16a-(lH-imidazol-l -yl)-pregnenolone acetate (2) from 16-DPA.
- Figure 2 shows the key NOESY correlations and relative stereochemistry of compounds 1 and 2. From this correlation the trarcs-orientation of the substituents in 16 and 17 positions was confirmed.
- the invented compounds l 6a-( l H-benzimidazol- l -yl)-pregnenolone acetate (1) and 16a-( l H-imidazol- l -yl)-pregnenolone acetate (2) are of pregnenolone acetate.
- One benzimidazole group and imidazole group are substituted at 16-position in compounds 1 and 2, respectively. These compounds were characterized by ⁇ NMR, l 3 C NMR and mass spectroscopy. These substituents at 16 position are a-oriented which are /ra -orientated with respect to the acetyl group at C- l 7 position.
- the present invention provides a process for the preparation of 16 «-( lH-benzimidazol-l -yl)-pregnenolone acetate (1) and 16oc-(lH-imidazol-l -yl)- pregnenolone acetate (2) by the solvent-free and catalyst-free Michael addition of nitrogen containing heterocycles benzimidazole and imidazole to 16- dehydropregnenolone acetate under microwave irradiation.
- the present invention also provides biological evaluation of compounds 1 and 2 as cytotoxic agent.
- Cellular viability in the presence of test compounds was determined by MTT- micro cultured tetrazolium assay.
- the cells seeded to flat bottom 96 ( l OOOcells/ ⁇ ⁇ ) well plates and cultured in the medium containing 1 0% serum and al lowed to attach and recover for 24 hours in a humid chamber containing 5% C0 2 MTT (3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide; sigma catalog No M2128) was dissolved in PBS at 5 mg/mL, filtered to sterilize and remove a small amount of insoluble residue present in MTT. Then different concentrations of the compound were added to the cells.
- compound 1 displayed good cytotoxicity to the tested cancer cell lines.
- Compound 1 had IC 50 values of 8.3317, 12.0192 and 8.2855 ⁇ against HeLa cell line, DU 145 cell line and MCF-7 cancer cell lines, respectively which is almost comparable to the drug doxorubicin.
- Compound 2 exhibited moderate activity against the tested cell lines. It had IC 50 values of 48.2584, 37.8069 and 20.0288 ⁇ against HeLa cell line, DU 145 cell line and MCF-7 cancer cell lines, respectively.
- the steroidal conjugated enone 16-DPA could be converted into corresponding Michael adduct 1 and 2 with benzimidazole and imidazole, respectively in high yield under microwave irradiation.
- the steroidal ketone 1 has shown cytotoxic activity similar to drug doxorubicin against I leLa cancer cell line, prostate DU 205 cancer cell line and breast cancer MCF-7 cell line.
- the crude product can be purified by neutral alumina also using EtOAc-hexane (1 :4) as the eluent to afford steroidal Michael adducts 1. (Yield 67%, 318 mg). Yellow solid, m.p.: 1 17 °C;
- the compound 16a-( l H-benzimidazol- l -yl)-pregnenolone acetate (1) exhibited cytotoxic activity against cervical HeLa cancer cell line, prostate DU 205 cancer cell line and breast cancer MCF-7 cell line similar to drug doxorubicin.
- the compound 16 ⁇ 3 ⁇ 4-( 1 H-imidazol- l -yl)-pregnenolone acetate (2) exhibited cytotoxic activity against cervical HeLa cancer cell l ine, prostate DU 205 cancer cell line and breast cancer MCF-7 cell line.
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Steroid Compounds (AREA)
Abstract
The present invention relates to microwave assisted facile Michael addition of benzimidazole and imidazole to steroidal α,β-unsaturated ketones under solvent-free condition to afford corresponding Michael adducts 16α-(1H-benzimidazol-1-yl)-pregnenolone acetate (1) and 16α-(1H-imidazol-1-yl)-pregnenolone acetate (2). Compound 1 showed in vitro cytotoxic activities almost comparable to the drug doxorubicin against cervical HeLa cancer cell line (compound 1, IC50 = 8.3317, doxorubicin IC50 = 7.8126), prostate DU 205 cancer cell line (compound 1, IC50 = 12.0192, doxorubicin IC50 = 9.1194) and breast cancer MCF-7 cell line (compound 1, IC50 = 8.2855, doxorubicin IC50 = 7.5094).
Description
16a-HETERO ARYL PREGNENOLONE ACETATE AND A PROCESS FOR
PREPARATION THEREOF
FIELD OF INVENTION
[0001] The present invention relates to 16a-heteroayl-l -yl pregnenolone acetate and a process for its preparation. The present invention particularly relates to compounds of 16cc-(lH-benzimidazol- l -yl)-pregnenolone acetate (1) and 16ct-(lH-imidazol- l -yl)- pregnenolone acetate (2). The present invention also relates to a process for the preparation of said compounds froml6-dehydropregnenolone acetate (16-DPA). More particularly the present invention relates to the anticancerous activity of the said compounds (1 and 2).
BACKGROUND OF THE INVENTION
[0002] Benzimidazole, imidazole and their derivatives are very important pharmacophores in medicinal chemistry showing a wide range of biological activities like antibacterial, antifungal, anticancer, antiviral, antituberculosis and antiinflammatory and antiageing agents.
[0003] Synthesis of several modified steroids comprising aza heterocyclic rings either fused or linked to the steroid framework have received considerable attention in the recent years due to their wide range of biological activities. These steroidal heterocycles are well known as inhibitors of enzymes such as 5 -reductase, 1 7 -Iyase and aromatase. Steroids comprising heterocyclic rings such as pyridine, imidazole, benzimidazole and diazine ring linked to 17-position show anticancer activity. Even 16-substituted steroids either by heterocycles or nonheterocyclic compounds have shown wide range of pharmacological activities. For example many \ 6E- arylidenosteroids are reported which show in vitro activity against many types of tumor cells. Even, 16-substituted steroids with imidazole heterocycle such as 16/5- imidazolyl androsterone derivatives are known which are synthesized using 17-oxo-5-
androsten-3 ?-ol as the starting material and these compounds are found to exhibit moderate cytotoxic effect in sixty cancer cell lines derived from nine cancer types.
[0004] Conjugated addition of benzimidazole/imidazole to a, ?-unsaturated carbonyl compounds are known using Lewis acid catalyzed addition, base catalyzed addition, high pressure promoted addition, iodine catalyzed addition, and acid catalyzed addition reactions. Unfortunately, many of these processes suffer from limitations, such as the use of expensive catalyst, expensive reagents, harsh conditions, relatively long reaction times, high catalyst loading, tedious work-up procedures and use of hazardous organic solvents. Hence, the development of a sustainable environment as an alternative procedure for Michael addition reaction avoiding the use of harmful solvents and expensive catalysts remains a challenging task to organic chemists. On the other hand, microwave-assisted organic synthesis (MAOS) which is recognized as a "green" technology has been applied as a very efficient way to accelerate the course of many organic reactions, producing high yields, higher selectivity, and lower quantities of side products.
[0005] Reference may be made to G A Potter, S E Barrie, M Jarman & M G Rowlands J. Med. Chem. 1995, 38, 2463 wherein, steroidal compounds having 17-(3- pyridyl) substituent together with 16, 17-double bond were synthesized which showed potent inhibition of human testicular 17«-hydroxylase-C i 7 2o-lyase. [0006] Another reference may be made to V D Handrattu, T S Vasaitis, V C O Njar, L K Gediya, R Kataria, P Chopra, D Newman, Jr.R Farquhar, Z Guo, Y Qi & A M H Brodie J. Med. Chem. 2005, 48, 2972 wherein, the syntheses and biological activities of novel 17-benzoazoles and 17-diazines were reported. The 17-benzimidazoIe steroidal compounds were shown to exhibit potent in vitro activities for CYP 17 inhibition, AR antagonism and inhibition of prostate cancer cell growth.
[0007] Still another reference may be made to R D Bruno, TS Vasaitis, L K Gediya, P Purnshottamachar, A M Godhole, Z Ates-Alagoz, A M H Brodie & V C O Njar
Steroids 2011, 76 1268 wherein it was found that 17-benzimidazole steroidal compound VN/ 124-1 was a potent inhibitor of human prostate cancer tumor growth and was remarkably more effective than castration or compound abiraterone that is currently undergoing phase III clinical trials in prostate cancer patients. [0008] Still another reference may be made to R Bansal & S Guleria Steroids 2008, 73, 1391 wherein the synthesis of 16-substituted steroidal compounds such as 16E-[3- methoxy-4-(2-aminoethoxy)-benzylidene]androstene derivatives and their cytotoxicity studies were reported.
[0009] Still another reference may be made to R Bansal, S Guleria, S Thota, S L Bodhankar, MR Patwardhan, C Zimmer, R W Hartmann & A L Harvey Steroids 2012, 77, 621 wherein, syntheses of 16 ?-imidazolyl-17-keto steroids and its 16 ?-imidazolyl- 17/?-hydroxyl derivatives were reported which exhibited moderate cytotoxic effects in sixty cancer cell lines derived from nine cancers types.
[0010] Still another reference may be made to J P Morrison, C A Stein, D S Casebier, J Morrison, C Stein and D Casebier WO/2011/017534^ wherein, compounds, pharmaceutical compositions and medicaments comprising such compounds, and methods of using such compounds that decrease androgen biosynthesis, decrease androgen receptor signaling and decrease androgen receptor sensitivity are reported.
[0011] Still another reference may be made to R Bansal, S Guleria, G Narang and H R Wolfgang WO/2007/031833, wherein a novel series of imidazolyl substituted steroidal and indan-l -one derivatives are reported.
[0012] Still another reference may be made to A Brodie and V C O Njar US5994335, wherein 17-azolyl steroids are reported to be useful as androgen synthesis inhibitors.
OBJECTIVES OF THE INVENTION
[0013] The main object of the present invention is to provide 16a-heteroayl- l -yl pregnenolone acetate and a process for preparation of the compounds.
[0014] Another object of the present invention is to provide compounds 16a-( lH- benzimidazol-l -yl)-pregnenolone acetate (1) and 16ec-(lH-imidazol-l -yl)- pregnenolone acetate (2).
[0015] Another object of the present invention is to provide a process for the preparation of 16a-(lH-benzimidazol-l -yl)-pregnenolone acetate (1) and 16a-(lH- imidazol-l -yl)-pregnenolone acetate (2) from 16-dehydropregnenolone acetate ( 16- DPA) without using solvent and catalyst under microwave irradiation.
[0016] Still another objective of the present invention is to evaluate the in vitro cytotoxic activities of compound 1 and 2 against cervical HeLa cancer cell line, prostate DU 205 cancer cell line and breast cancer MCF-7 cell line in comparison to the drug doxorubicin.
SUMMARY OF INVENTION
[0017] Accordingly the present invention provides a compound of formula A,
[0018] In an embodiment of the present invention compound, wherein structural formula of representative compound comprising are:
1 ( C30H 38 N 2O 3 ) 2 (C 26 H36 N203 )
[0019] In another embodiment of the present invention a compound wherein the formula is represented by the group of the following compounds:- a) 16a-(lH-benzimidazol-l -yl)-pregnenolone acetate (1)
b) 16a-(lH-imidazol-l -yl)-pregnenolone acetate (2)
[0020] In another embodiment of the present invention a compound wherein 16a- (lH-benzimidazol-l -yl)-pregnenolone acetate (1) and 16a-(lH-imidazol-l-yl)- pregnenolone acetate (2) exhibited in vitro cytotoxic activity against cervical HeLa cancer cell line, prostate DU 205 cancer cell line and breast cancer MCF-7 cell line.
[0021] In another embodiment of the present invention a compound wherein cytotoxic activities of compound 1 with ICso= 8.3317 against HeLa cancer cell line, IC5o= 12.0192 against prostate DU 205 cancer cell line and IC50= 8.2855 against breast cancer MCF-7 cell line.
[0022] In yet another embodiment of the present invention a compound wherein cytotoxic activities of compound 2 with IC50= 48.2584 against HeLa cancer cell line, IC50= 37.8069 against prostate DU 205 cancer cell line and IC50= 20.0288 against breast cancer MCF-7 cell line.
[0023] In yet another embodiment of the present invention a process for the preparation of compound of formula ( I ) wherein the said process comprises of following steps:- a) mixing a, ?-unsaturated ketone 16-DPA with compounds selected from benzimidazole or imidazole in mortar,
b) irradiating the mixture obtained in step a) in a microwave reactor at a frequency of 2450 MHz for a time period ranging between 10 to 30 minutes,
c) purifying the crude product obtained in step b) by silica gel column chromatography using EtOAc-hexane taken in the ratio of 1 :4.
[0024] In another embodiment of the present invention a process wherein the compounds are prepared by solvent free and catalyst-free method by the Michael addition of benzimidazole to 16-DPA under microwave irradiation.
[0025] In another embodiment of the present invention a process, wherein the yield of the compound of formula A is as high as 47% to 86%.
BRIEF DESCRIPTION OF DRAWINGS & FIGURES
[0026] Figure 1 shows the synthesis of compounds 16a-(lH-benzimidazol-l -yl)- pregnenolone acetate (1) and 16a-(lH-imidazol-l -yl)-pregnenolone acetate (2) from 16-DPA. [0027] Figure 2 shows the key NOESY correlations and relative stereochemistry of compounds 1 and 2. From this correlation the trarcs-orientation of the substituents in 16 and 17 positions was confirmed.
DETAILED DESCRIPTION OF THE INVENTION
1 ( C30H38N2O3) 2 (C2BH36N203)
[0028] The invented compounds l 6a-( l H-benzimidazol- l -yl)-pregnenolone acetate (1) and 16a-( l H-imidazol- l -yl)-pregnenolone acetate (2) are of pregnenolone acetate. One benzimidazole group and imidazole group are substituted at 16-position in compounds 1 and 2, respectively. These compounds were characterized by Ή NMR, l 3C NMR and mass spectroscopy. These substituents at 16 position are a-oriented which are /ra -orientated with respect to the acetyl group at C- l 7 position. This /ram- orientation is proved by the NOESY spectra of both the compounds 1 and 2. Compound 1 is a yellow solid, (m.p.: 1 1 8 °C) and compound 2 is a brown solid (m.p.:
131 °C). Both the compounds 1 and 2 show in vitro cytotoxic activity against cervical HeLa cancer cell line, prostate DU 205 cancer cell line and breast cancer MCF-7 cell line.
[0029] Accordingly, the present invention provides a process for the preparation of 16«-( lH-benzimidazol-l -yl)-pregnenolone acetate (1) and 16oc-(lH-imidazol-l -yl)- pregnenolone acetate (2) by the solvent-free and catalyst-free Michael addition of nitrogen containing heterocycles benzimidazole and imidazole to 16- dehydropregnenolone acetate under microwave irradiation. The present invention also provides biological evaluation of compounds 1 and 2 as cytotoxic agent. [0030] The reaction of equimolar amount of 16-DPA (1) and benzimidazole in protic solvents ethanol and isopropanol under thermal heating conditions could not provide the product 1 (Figure 1 ). Similar results were obtained using nonprotic solvents acetonitrile and dimethylformamide under reflux condition (8 hours). Interestingly, when the reaction was performed in a solvent-free condition, under thermal heating ( 140 °C) for 8 hours, the product 1 was obtained in satisfactory yield (40%). It was noteworthy to observe that finely ground equimolar amounts of 1 and benzimidazole under microwave irradiation (720 Watt) for 10 minutes afforded 1 in 69% yield. The formation of 16et-substituted N-heterocycles having trans orientation with respect to the acetyl group at C- 17 position of the steroid moiety was determined by comparing the optical rotational data of these hetero steroids with the reported 16-substituted steroidal heterocycles by Gould et al. (D Gould, E L Shapiro, L E Finckenor, F Gruen & E B Hershberg J. Am. Chem. Soc. 1956, 78, 3158). The a orientation of benzimidazole at 16-position and the β orientation of acetyl group at 17-position was then confirmed by NOESY spectrum of compound 1. In the NOESY spectrum of compound 1, the presence of correlation of CH3-18 protons with H- 16 and CH3- I 8 protons with H-21 protons and the absence of correlation between CH3- I 8 protons with H- 1 7 also indicated that H- 16 and H- 17 were β and a oriented respectively in compound 1 (Figure 2). Similarly, the reaction of 16-DPA with imidazole under
microwave irradiation (720 Watt) for 10 minutes afforded Michael adduct 2 in 86% yield under microwave irradiation (Figure 1 ).
16-DPA
Figure 1 [0031] In vitro cytotoxic activity studies of 16a-( lH-benzimidazol-l -yl)- pregnenolone acetate (1):
[0032] These cell lines were procured from ATCC in 2009. Catalog numbers HeLa (ATCC®CCL-2TM)-15th passage cells used for the present experiment. DU 145 (ATCC® HTB-81™)-12 the passage cells used for the present experiment. MCF7 (ATCC® HTB-22™)-l 5 the passage cells used for the present experiment.
[0033] Cellular viability in the presence of test compounds was determined by MTT- micro cultured tetrazolium assay. The cells seeded to flat bottom 96 ( l OOOcells/Ι ΟΟμΙ) well plates and cultured in the medium containing 1 0% serum and al lowed to attach and recover for 24 hours in a humid chamber containing 5% C02 MTT (3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide; sigma catalog No M2128) was dissolved in PBS at 5 mg/mL, filtered to sterilize and remove a small amount of insoluble residue present in MTT. Then different concentrations of the compound were added to the cells. After 48 hours, stock MTT solution ( 10 μΐ) was added to the culture plate. Cells were again kept in C02 incubator for 2 hours. After incubation, 100 μΙ of DMSO was added and mixed. The absorbance was read at 562 nm in a plate reader. The results were represented as percentage of cytotoxicity/viability. All the experiments were carried out in triplicates. From the percentage of cytotoxicity the 1C-
50 value was calculated (S Myadaraboina, M Alia, V Saddanapu, V R Boenaa & A Addlagatta. Eur. J. Med. Chem. 2010, 45, 5208).
[0034] As showed in Table 1 , compound 1 displayed good cytotoxicity to the tested cancer cell lines. Compound 1 had IC50 values of 8.3317, 12.0192 and 8.2855 μΜ against HeLa cell line, DU 145 cell line and MCF-7 cancer cell lines, respectively which is almost comparable to the drug doxorubicin. Compound 2 exhibited moderate activity against the tested cell lines. It had IC50 values of 48.2584, 37.8069 and 20.0288 μΜ against HeLa cell line, DU 145 cell line and MCF-7 cancer cell lines, respectively.
[0035] The steroidal conjugated enone 16-DPA could be converted into corresponding Michael adduct 1 and 2 with benzimidazole and imidazole, respectively in high yield under microwave irradiation. The steroidal ketone 1 has shown cytotoxic activity similar to drug doxorubicin against I leLa cancer cell line, prostate DU 205 cancer cell line and breast cancer MCF-7 cell line.
EXAMPLES
[0036] The following examples are given by illustrations and should not construe the scope of the invention.
Example-I
[0037] Preparation of 16ct-( 1 H-benzimidazol- 1 -yl)-pregnenolone acetate (1):
α, Jnsaturated ketone 16-DPA (356 mg, 1 .0 mmol) and benzimidazole (1 18 mg, 1 .0 mmol) were mixed intimately in a mortar and irradiated in a microwave reactor (720 Watt, 2450 MHz) for 10 minutes. On completion of the reaction (by TLC), the crude product was purified by silica gel column chromatography using EtOAc-hexane (1 :4) as the eluent to afford steroidal Michael adduct 1. (Yield 69%, 327 mg). The crude product can be purified by neutral alumina also using EtOAc-hexane (1 :4) as the eluent to afford steroidal Michael adducts 1. (Yield 67%, 318 mg). Yellow solid, m.p.: 1 17 °C;
Rf = 0.3 (EtOAc/Hexane = 1 :4); [a]D = -6.6 (c, 0.5, CHC13); IR (CHC13, cm-1): 2927, 2856, 1732, 1709, 1655, 1361, 1250, 1033, 762; Ή NMR (CDC13, 300 MHz): δ 8.10 (s, 1H), 7.79 (dd, 1H, J = 6.8 & 3.5 Hz), 7.50 (dd, 1H, J = 8.2 & 2.0 Hz), 7.33-7.23 (m, 2H), 5.59-5.53 (m, 1H), 5.41-5.38 (m, 1H), 4.66-4.60 (m, 1H), 3.06 (d, 1H, J = 8.5 Hz), 2.09 (s, 3H), 2.05 (s, 3H), 1.07 (s, 3H), 2.36-0.89 (m, 17H), 0.81 (s, 3H); 13C NMR (CDC13, 75 MHz): δ 206.2, 170.6, 144.0, 142.1 , 139.9, 132.7, 122.9, 122.2, 121.7, 120.5, 1 10.5, 73.7, 70.6, 58.4, 56.6, 54.6, 49.7, 45.2, 38.6, 38.0, 36.9, 36.6, 32.0, 31 .6, 27.7, 21 .4, 20.7, 19.3, 18.4, 13.9; MS (EI, m/z) = 474 (M+), 414 (M+-60). Anal, calcd. for C30H38 2O3 : C, 75.92; H, 8.07; N, 5.90. Found: C, 76.22; H, 8.18; N, 6.03.
[0038] Preparation of 3p-acetoxy- 16a-( l H-imidazol-l -yl)-17p-acetyl-androst-5-ene
16 DPA (356 mg, 1 .0 mmol) and imidazole (68 mg, 1.0 mmol) were mixed intimately in a mortar and irradiated in a microwave reactor (720 Watt) for 10 minutes. On completion of the reaction (by TLC), the crude product was purified by silica gel column chromatography using EtOAc-hexane ( 1 :4) as the eluent to afford steroidal Michael adduct 2. Brown solid, m.p.: 130 °C. Yield 86% (365 mg); Rr: 0.4 (EtOAc/Hexane = 3:7); [a]D = -8.4 (c 0.5, CHCI3); IR (CHCI3) cm"' : v 2943, 2854, 1731 , 1705, 1649, 1 364, 1247, 1034, 754; Ή NMR (CDC13, 300 MHz): δ 7.55 (s, 1 H), 7.03 (s, 1 H), 6.91 (s, 1 H), 5.40-5.37 (m, 1 H), 5.30-5.22 (m, 1 H), 4.67-4.57 (m, 1 H), 2.80 (d, 1 H, J = 7.8 Hz), 2.10 (s, 3H), 2.04 (s, 3H), 1 .04 (s, 3H), 0.73 (s, 3H), 2.40-0.89 (m, 1 7H); l 3C NMR (CDC13, 75 MHz): δ 206.0, 170.6, 141 .4, 139.8 (2C), 121 .7 (2C),
73.7, 72.8, 56.1 , 55.8, 49.5, 45.2, 38.0, 36.6, 33.6, 31.9, 31.5, 29.7, 29.4, 27.7, 22.7, 21.4, 20.6, 19.3, 14.1 ; MS (EI) m/z = 364 (M+-60), 296 [(M+-60)-imidazole]. Anal, calcd. for C26H36N203: C, 73.55; H, 8.55; N, 6.60; Found: C, 73.69; H, 8.51 ; N, 6.80. Example-II
[0039] Preparation of 16a-(lH-benzimidazol-l -yl -pregnenolone acetate (1): a, ?-Unsaturated ketone 16-DPA (2.136 g, 6.0 mmol) and benzimidazole (708 mg, 6.0 mmol) were mixed intimately in a mortar and irradiated in a microwave reactor (720 Watt, 2450 MHz) for 10 minutes. On completion of the reaction (by TLC), the crude product was purified by silica gel column chromatography using EtOAc-hexane (1 :4) as the eluent to afford steroidal Michael adduct 1. Yellow solid, m.p.: 1 19 °C. Yield 68% (1.93 g).
[0040] Preparation of 3P-acetoxy-16a-(l H-irnidazol-l -yl)-17p-acetyl-androst-5-ene £2)1
16 DPA (2.136 g, 6.0 mmol) and imidazole (408 mg, 6.0 mmol) were mixed intimately in a mortar and irradiated in a microwave reactor (720 Watt) for 10 minutes. On completion of the reaction (by TLC), the crude product was purified by silica gel column chromatography using EtOAc-hexane ( 1 :4) as the eluent to afford steroidal Michael adduct 2. Brown solid, m.p.: 13 1 °C. Yield 85% (2.16 g).
Example-Ill
[0041] Preparation of 16a-( l H-benzimidazol-l -yl)-pregnenolone acetate (1 ): a, ?-Unsaturated ketone 16-DPA (4.272 g, 12.0 mmol) and benzimidazole ( 1 .42 g, 12.0 mmol) were mixed intimately in a mortar and irradiated in a microwave reactor (720 Watt, 2450 MHz) for 10 minutes. On completion of the reaction (by TLC), the crude product was purified by silica gel column chromatography using EtOAc-hexane ( 1 :4) as the eluent to afford steroidal Michael adduct 1. Yellow solid, m.p.: 1 18 °C. Yield 69% (3.92 g).
[0042] Preparation of 3 β-acetoxy- 16 -( 1 H-imidazol- 1 -yl)- 1 7P-acetyl-androst-5-ene
16 DPA (4.272 g, 12.0 mmol) and imidazole (816 mg, 12.0 mmol) were mixed intimately in a mortar and irradiated in a microwave reactor (720 Watt, 2450 MHz) for 10 minutes. On completion of the reaction (by TLC), the crude product was purified by silica gel column chromatography using EtOAc-hexane (1 :4) as the eiuent to afford steroidal Michael adduct 2. Brown solid, m.p.: 130 °C. Yield 83% (4.22 g).
Example-IV
[0043] Preparation of 16a-( l H-benzimidazol-l -yr)-pregnenolone acetate (1): a, ?-Unsaturated ketone 16-DPA (356 mg, 1.0 mmol) and benzimidazole (1 18 mg, 1.0 mmol) were mixed intimately in a mortar and irradiated in a microwave reactor (540 Watt, 2450 MHz) for 10 minutes. On completion of the reaction (by TLC), the crude product was purified by silica gel column chromatography using EtOAc-hexane ( 1 :4) as the eiuent to afford steroidal Michael adduct 1. (Yield 47%, 223 mg). Yellow solid, m.p.: 1 17 °C.
[0044] Preparation of 3p-acetoxy-16 -(l H-imidazol-l -yl)-17P-acetyl-androst-5-ene £2):
16 DPA (356 mg, 1 .0 mmol) and imidazole (68 mg, 1 .0 mmol) were mixed intimately in a mortar and irradiated in a microwave reactor (540 Watt, 2450 MHz) for 10 minutes. On completion of the reaction (by TLC), the crude product was purified by silica gel column chromatography using EtOAc-hexane ( 1 :4) as the eiuent to afford steroidal Michael adduct 2. Brown solid, m.p.: 1 30 °C. Yield 62% (263 mg).
Example-V
[0045] Preparation of 16a-( l H-benzimidazol- l -yl)-pregnenolone acetate (1): a,/?-Unsaturated ketone 16-DPA (356 mg, 1.0 mmol) and benzimidazole ( 1 1 8 mg, 1 .0 mmol) were mixed intimately in a mortar and irradiated in a microwave reactor (540 Watt, 2450 MHz) for 20 minutes. On completion of the reaction (by TLC), the crude product was purified by silica gel column chromatography using EtOAc-hexane ( 1 :4) as the eiuent to afford steroidal Michael adduct 1. (Yield 52%, 247 mg). Yellow solid, m.p.: 1 1 7 °C.
[0046] Preparation of 3 β-acetoxy- 16a-( 1 H-imidazol- 1 -vD- 17p-acetyl-androst-5-ene £211
16 DPA (356 mg, 1.0 mmol) and imidazole (68 mg, 1.0 mmol) were mixed intimately in a mortar and irradiated in a microwave reactor (540 Watt, 2450 MHz) for 20 minutes. On completion of the reaction (by TLC), the crude product was purified by silica gel column chromatography using EtOAc-hexane (1 :4) as the eluent to afford steroidal Michael adduct 2. Brown solid, m.p.: 130 °C. Yield 66% (280 mg).
Example- VI
[0047] Preparation of 16a-(lH-benzimidazol-l-yl)-pregnenolone acetate (1): α, Jnsaturated ketone 16-DPA (356 mg, 1.0 mmol) and benzimidazole (1 18 mg, 1.0 mmol) were mixed intimately in a mortar and irradiated in a microwave reactor (720 Watt, 2450 MHz) for 20 minutes. On completion of the reaction (by TLC), the crude product was purified by silica gel column chromatography using EtOAc-hexane (1 :4) as the eluent to afford steroidal Michael adduct 1. (Yield 69%, 327 mg). Yellow solid, m.p.: 1 16 °C.
[0048] Preparation of 3 β-acetoxy- 16a-( 1 H-imidazol- 1 -yl)- 17P-acetyl-androst-5-ene
16 DPA (356 mg, 1 .0 mmol) and imidazole (68 mg, 1 .0 mmol) were mixed intimately in a mortar and irradiated in a microwave reactor (720 Watt, 2450 MHz) for 20 minutes. On completion of the reaction (by TLC), the crude product was purified by silica gel column chromatography using EtOAc-hexane ( 1 :4) as the eluent to afford steroidal Michael adduct 2. Brown solid, m.p.: 131 °C. Yield 86% (365 mg).
Example-VII
10049] Preparation of 16a-( l H-benzimidazol- l -yl)-pregnenolone acetate (1): α,/3-Unsaturated ketone 16-DPA (356 mg, 1.0 mmol) and benzimidazole ( 1 1 8 mg, 1 .0 mmol) were mixed intimately in a mortar and irradiated in a microwave reactor (900 Watt, 2450 MHz) for 10 minutes. On completion of the reaction (by TLC), the crude
product was purified by silica gel column chromatography using EtOAc-hexane (1 :4) as the eluent to afford steroidal Michael adduct 1. (Yield 65%, 308 mg). Yellow solid, m.p.: 1 16 °C.
[0050] Preparation of 3p-acetoxy- 16a-( 1 H-imidazol- 1 -yl)- 17P-acetyl-androst-5-ene (21:
16 DPA (356 mg, 1.0 mmol) and imidazole (68 mg, 1.0 mmol) were mixed intimately in a mortar and irradiated in a microwave reactor (900 Watt, 2450 MHz) for 10 minutes. On completion of the reaction (by TLC), the crude product was purified by silica gel column chromatography using EtOAc-hexane ( 1 :4) as the eluent to afford steroidal Michael adduct 2. Brown solid, m.p.: 132 °C. Yield: 79% (335 mg).
ADVANTAGES OF THE PRESENT INVENTION [0051] The main advantages of the present invention are:
1 . The compound 16a-( l H-benzimidazol- l -yl)-pregnenolone acetate (1) exhibited cytotoxic activity against cervical HeLa cancer cell line, prostate DU 205 cancer cell line and breast cancer MCF-7 cell line similar to drug doxorubicin.
2. The compound 16<¾-( 1 H-imidazol- l -yl)-pregnenolone acetate (2) exhibited cytotoxic activity against cervical HeLa cancer cell l ine, prostate DU 205 cancer cell line and breast cancer MCF-7 cell line.
3. Incorporation of a benzimidazole group and imidazole group at 16-position of steroidal D-ring via solvent and catalyst free Michael addition reaction of benzimidazole and imidazole with commercially available 16-DPA molecule under microwave irradiation.
4. Incorporation of a benzimidazole/imidazole group at 16-position affords 16a- substituted heterosteroid which has /ram-orientation with respect to 17-acetyl group.
Claims
1. A compound of formula A,
1 (C30H38N2O3) 2 (C26H36N203)
3. A compound as claimed in claim 1 wherein the formula is represented by the group of the following compounds:- a) 16a-(lH-benzimidazol-l-yl)-pregnenolone acetate (1)
b) 16a-(l H-imidazol-l-yl)-pregnenolone acetate (2)
4. A compound as claimed in claim 1 wherein 16 -(lH-benzimidazol-l-yl)- pregnenolone acetate (1) and 16a-(lH-imidazol-l-yl)-pregnenolone acetate (2) exhibited in vitro cytotoxic activity against cervical HeLa cancer cell line, prostate DU 205 cancer cell line and breast cancer MCF-7 cell line.
A compound as claimed in claim 1 wherein cytotoxic activities of compound 1 with 1C50= 8.3317 against HeLa cancer cell line, IC5o= 12.0192 against prostate DU 205 cancer cell line and IC5o= 8.2855 against breast cancer MCF-7 cell line. A compound as claimed in claim 1 wherein cytotoxic activities of compound 2 with IC5o= 48.2584 against HeLa cancer cell line, IC50= 37.8069 against prostate DU 205 cancer cell line and lC5o= 20.0288 against breast cancer MCF- 7 cell line.
A process for the preparation of compound of formula (1) wherein the said process comprises of following steps:- a) mixing a, ?-unsaturated ketone 16-DPA with compounds selected from benzimidazole or imidazole in mortar,
b) irradiating the mixture obtained in step a) in a microwave reactor at a frequency of 2450 MHz for a time period ranging between 10 to 30 minutes,
c) purifying the crude product obtained in step b) by silica gel column chromatography using EtOAc-hexane taken in the ratio of 1 :4.
A process as claimed in claim 7 wherein the compounds are prepared by solvent free and catalyst-free method by the Michael addition of benzimidazole to 16- DPA under microwave irradiation.
A process as claimed in claim 7, wherein the yield of compound of formula A is as high as 47% to 86%.
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| CN113831381A (en) * | 2020-06-24 | 2021-12-24 | 中国科学院上海药物研究所 | Steroid compound for treating depression |
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| CN113831382A (en) * | 2020-06-24 | 2021-12-24 | 中国科学院上海药物研究所 | Steroid compound |
| CN113831381A (en) * | 2020-06-24 | 2021-12-24 | 中国科学院上海药物研究所 | Steroid compound for treating depression |
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